High amplitude stretching of ATII cells and fibroblasts results in profibrotic effects

Abstract

Background: Inappropriate mechanical forces act on alveolar epithelial cells during mechanical ventilation e.g. in ARDS and possibly in patients with pulmonary fibrosis. These forces can cause lung injury and may contribute to the development or aggravation of pulmonary fibrosis. Aim of the study: We investigated the hypothesis that high amplitude mechanical stretching of alveolar type II (ATII) cells and lung fibroblasts promotes profibrotic processes. Material and Methods: ATII cells and fibroblasts were stretched on elastic membranes using a pattern of higher amplitudes (“unphysiological”). The production of profibrotic cytokines and extra cellular matrix (ECM) proteins were investigated in supernatants. In addition, we determined the expression of relevant microRNAs (miRNA) and the process of epithelial-mesenchymal transition (EMT) in ATII cells. Results: Unphysiological stretch of ATII cells led to increased release of TGF-β1 into supernatants. We also found elevated protein levels of collagen I and IV in supernatants of stretched cells. By contrast, stretching of fibroblasts changed neither the expression of fibrosis-modulating factors nor ECM-proteins. However, fibroblasts significantly withstood stretch-induced cell injury and seemed to have a survival benefit. Further, stretched ATII cells exhibited a higher expression of miRNAs (miR-15b, miR-25, let-7d) relevant to EMT. The process of EMT, which is characterized by an increase of vimentin and a decrease of cytokeratin expression, was significantly accelerated due to stretching of ATII cells. Conclusion: These data provide evidence that unphysiological mechanical stretching of lung cells induced several profibrotic effects and accelerated EMT, which may have critical implications in terms of development or aggravation of pulmonary fibrosis in the clinical context.

Keywords:

• Apoptosis
• EMT
• fibrosis
• lung injury
• stretching

Acknowledgment

We thank Claudia Haenel for her excellent technical support.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Additional information

Funding

Dr Armin Frille was temporarily supported by the Federal Ministry of Education and Research (BMBF), Germany, FKZ: 01EO1501 (IFB Adiposity Diseases, postdoctoral fellowship ‘MetaRot program’).